• Title/Summary/Keyword: Multiple enzymes

Search Result 167, Processing Time 0.026 seconds

A microfluidic multiple bio-analysis platform based on the enzyme-immobilized barcoded strip

  • Kim, Sung-Rak;Lee, Sang-Hoon
    • 한국생물공학회:학술대회논문집
    • /
    • 2005.10a
    • /
    • pp.159-162
    • /
    • 2005
  • In this paper, we report a novel technique for the manufacture of polymeric bar-coded strips having diverse characteristics such as sensing with biocatalysts using a microfluidic platform and 'on the fly' photopolymerization. This method is a very simple, cost-effective means for mass production, and diverse materials sensitive to hazardous environments such as enzymes, DNA, or antigens are expected to be immobilized stably, as the fabrication process does not need any hazardous environments. On the basis of this technology, we fabricated enzyme-immobilized barcoded strip for multiple bio-analysis.

  • PDF

Evaluation of Adverse Drug Properties with Cryopreserved Human Hepatocytes and the Integrated Discrete Multiple Organ Co-culture (IdMOCTM) System

  • Li, Albert P.
    • Toxicological Research
    • /
    • v.31 no.2
    • /
    • pp.137-149
    • /
    • 2015
  • Human hepatocytes, with complete hepatic metabolizing enzymes, transporters and cofactors, represent the gold standard for in vitro evaluation of drug metabolism, drug-drug interactions, and hepatotoxicity. Successful cryopreservation of human hepatocytes enables this experimental system to be used routinely. The use of human hepatocytes to evaluate two major adverse drug properties: drug-drug interactions and hepatotoxicity, are summarized in this review. The application of human hepatocytes in metabolism-based drug-drug interaction includes metabolite profiling, pathway identification, P450 inhibition, P450 induction, and uptake and efflux transporter inhibition. The application of human hepatocytes in toxicity evaluation includes in vitro hepatotoxicity and metabolism-based drug toxicity determination. A novel system, the Integrated Discrete Multiple Organ Co-culture (IdMOC) which allows the evaluation of nonhepatic toxicity in the presence of hepatic metabolism, is described.

BIOCHEMICAL CHARACTERISTICS OF SALIVA TO BE LINKED TO THE MULTIPLE CARIES IN CHILDREN (소아의 다발성 치아우식증과 연관된 타액의 생화학적 특성)

  • Chang, Hee-Soon;Cho, Woo-Sung;Choi, Byung-Jai;Seo, Jeong-Taeg;Lee, Syng-II
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.25 no.4
    • /
    • pp.691-703
    • /
    • 1998
  • Saliva is obviously potential medium to protect the dental caries by not only physical clearing effect, but aggregating action of protein with bacteria. Nevertheless, we still do not understand how the dental caries occur and what brings the individual difference in caries prevalence. In the regards of dental caries prevalence, we hypothesized that the composition of salivary protein might be different from caries susceptible group to caries resistant group. The purposes of this experiment were focused on the molecular analysis of salivary proteins from the subjects who were involved in multiple caries. Electrophoretic analysis was done on the whole saliva collected from the children with and without multiple caries. We found 86.2% of subjects with multiple caries has approximately 120 KDa protein band while 30.4% in the healthy subjects. And the concentration of the total protein on the subjects with multiple caries is significantly higher than that of the healthy group. However, it turned out that the difference of the salivary composition does not affect the bacterial adhesion to hydroxyapatite bead. With regards of enzymes in saliva, the activity of ${\alpha}-amylase$ and lactate dehydrogenase does not have any significant difference between both groups. However, the concentrations of $Na^+\;and\;Cl^-$ in saliva from multiple caries group is higher than that of the control group. Taken all together, it may be concluded that 120 KDa protein in saliva may be associated with the process of dental caries, also the high concentration of protein and $Na^+,\;Cl^-$ in saliva may be linked to dental caries development as a cofactors.

  • PDF

Production of Endoglucanase, Beta-glucosidase and Xylanase by Bacillus licheniformis Grown on Minimal Nutrient Medium Containing Agriculture Residues

  • Seo, J.;Park, T.S.;Kim, J.N.;Ha, Jong K.;Seo, S.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.27 no.7
    • /
    • pp.946-950
    • /
    • 2014
  • Bacillus licheniformis was grown in minimal nutrient medium containing 1% (w/v) of distillers dried grain with soluble (DDGS), palm kernel meal (PKM), wheat bran (WB) or copra meal (CM), and the enzyme activity of endoglucanase, ${\beta}$-glucosidase, xylanase and reducing sugars was measured to investigate a possibility of using cost-effective agricultural residues in producing cellulolytic and hemicellulolytic enzymes. The CM gave the highest endoglucanase activity of 0.68 units/mL among added substrates at 48 h. CM yielded the highest titres of 0.58 units/ml of ${\beta}$-glucosidase, compared to 0.33, 0.23, and 0.16 units/mL by PKM, WB, and DDGS, respectively, at 72 h. Xylanase production was the highest (0.34 units/mL) when CM was added. The supernatant from fermentation of CM had the highest reducing sugars than other additional substrates at all intervals (0.10, 0.12, 0.10, and 0.11 mg/mL respectively). It is concluded that Bacillus licheniformis is capable of producing multiple cellulo- and hemicellololytic enzymes for bioethanol production using cost-effective agricultural residues, especially CM, as a sole nutrient source.

Effect of 6-Aminonicotinamide on the Levels of Some Metabolites and Related Enzymes in Rabbit Serum (6-Aminonicotinamide가 토끼혈청내 효소 및 대사물질에 미치는 영향)

  • Park, In-Koo;Lee, Chul-Seung;Lee, Seung-Hoon;Song, Yoon-Kyung;Shin, Sook
    • The Korean Journal of Zoology
    • /
    • v.33 no.4
    • /
    • pp.493-498
    • /
    • 1990
  • The effects of an antimetabolite, 6-aminonicotinamide (6-AN) on the levels of enzymes and metabolites in rabbit serum were investigated. The intraperitoneal administration of 6-AN (multiple doses of l5mg/kg body weight) gave tise to a remarkable increase in glucose and cholesterol levels but did not exert any appreciable influence on the concentration of albumin and total protein. Alkaline phosphatase activity was significantly reduced by administration of 6-AN, whereas creatine phophokinase, serum glutamic oxaloacetate transaminase and serum glutamic pyruvate transaminase activities were matkedly enhanced. Nevettheless, the levels of Ca, P, Na, K, Cl and Co were not affeded to any extent by 6-AN.

  • PDF

Screening and Characterization of Psychrotrophic, Lipolytic Bacteria from Deep-Sea Sediments

  • Zeng, Xiang;Xiao, Xiang;Wang, Peng;Wang, Rengping
    • Journal of Microbiology and Biotechnology
    • /
    • v.14 no.5
    • /
    • pp.952-958
    • /
    • 2004
  • Of 23 psychrotrophic bacteria isolated from the west Pacific deep-sea sediments, 19 were assigned to the $\gamma$-Proteobacteria, 3 to the <$\beta$-Proteobacteria, and 1 to the Gram-positive bacteria, as determined by their 16S rDNA sequences. Ten psychrotrophs, affiliated to the Psychrobacter, Pseudoalteromonas, and Pseudomonas genera in the $\gamma$-Proteobacteria group, were screened for lipolytic bacteria. The majority of the lipolytic isolates had growth temperatures between 4-$30^\circ{C}$, and all of them were neutrophilic, aerobic, or facultatively anaerobic, and some were able to produce multiple kinds of ectohydrolytic enzymes. The deep-sea strains Psychrobacter sp. wp37 and Pseudoalteromonas sp. wp27 were chosen for further lipase production analysis. Both strains had the highest lipase production when grown at 10 to $20^\circ{C}$; their highest lipase production occurred at the late-exponential growth stage; and the majority of the enzymes were excreted to the outside of the cells. Lipases from both strains had the same optimal reaction temperature and pH (20-$30^\circ{C}$, pH 7-8) and could retain about 60% of their highest activity at $4^\circ{C}$. Furthermore, SDS-PAGE and an in-gel activity test showed that they had the same high molecular mass of about 85 kDa.

Expression of Catalase (CAT) and Ascorbate Peroxidase (APX) in MuSI Transgenic Tobacco under Cadmium Stress

  • Kim, Kye-Hoon;Kim, Young-Nam;Lim, Ga-Hee;Lee, Mi-Na;Jung, Yoon-Hwa
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.44 no.1
    • /
    • pp.53-57
    • /
    • 2011
  • The MuSI is known as a multiple stress resistant gene with several lines. A previous study using RT-PCR showed that the expression of MuSI gene in tobacco plant induced its tolerance to Cd stress. This study was conducted to examine the enhanced Cd tolerance of the MuSI transgenic tobacco plant through germination test and to understand the role of the involved antioxidant enzymes for the exhibited tolerance. Germination rate of MuSI transgenic tobacco was more than 10% higher than that of wild-type tobacco, and seedlings of MuSI transgenic tobacco grew up to 1.6 times larger and greener than seedlings of wild-type tobacco at 200 and 300 ${\mu}M$ Cd. From the third to the fifth day, CAT activities at 100 and 200 ${\mu}M$ Cd and APX activities at 100, 200 and 300 ${\mu}M$ Cd of MuSI transgenic tobacco were up to two times higher than those of wild-type tobacco. MuSI gene is shown to enhance the activities of antioxidant enzymes resulting in higher tolerance to oxidative stress compared with the control plant.

Enzymatic DNA oxidation: mechanisms and biological significance

  • Xu, Guo-Liang;Walsh, Colum P.
    • BMB Reports
    • /
    • v.47 no.11
    • /
    • pp.609-618
    • /
    • 2014
  • DNA methylation at cytosines (5mC) is a major epigenetic modification involved in the regulation of multiple biological processes in mammals. How methylation is reversed was until recently poorly understood. The family of dioxygenases commonly known as Ten-eleven translocation (Tet) proteins are responsible for the oxidation of 5mC into three new forms, 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). Current models link Tet-mediated 5mC oxidation with active DNA demethylation. The higher oxidation products (5fC and 5caC) are recognized and excised by the DNA glycosylase TDG via the base excision repair pathway. Like DNA methyltransferases, Tet enzymes are important for embryonic development. We will examine the mechanism and biological significance of Tet-mediated 5mC oxidation in the context of pronuclear DNA demethylation in mouse early embryos. In contrast to its role in active demethylation in the germ cells and early embryo, a number of lines of evidence suggest that the intragenic 5hmC present in brain may act as a stable mark instead. This short review explores mechanistic aspects of TET oxidation activity, the impact Tet enzymes have on epigenome organization and their contribution to the regulation of early embryonic and neuronal development.

Tetrahydropteridines possess antioxidant roles to guard against glucose-induced oxidative stress in Dictyostelium discoideum

  • Park, Seon-Ok;Kim, Hye-Lim;Lee, Soo-Woong;Park, Young Shik
    • BMB Reports
    • /
    • v.46 no.2
    • /
    • pp.86-91
    • /
    • 2013
  • Glucose effects on the vegetative growth of Dictyostelium discoideum Ax2 were studied by examining oxidative stress and tetrahydropteridine synthesis in cells cultured with different concentrations (0.5X, 7.7 g $L^{-1}$; 1X, 15.4 g $L^{-1}$; 2X, 30.8 g $L^{-1}$) of glucose. The growth rate was optimal in 1X cells (cells grown in 1X glucose) but was impaired drastically in 2X cells, below the level of 0.5X cells. There were glucose-dependent increases in reactive oxygen species (ROS) levels and mitochondrial dysfunction in parallel with the mRNA copy numbers of the enzymes catalyzing tetrahydropteridine synthesis and regeneration. On the other hand, both the specific activities of the enzymes and tetrahydropteridine levels in 2X cells were lower than those in 1X cells, but were higher than those in 0.5X cells. Given the antioxidant function of tetrahydropteridines and both the beneficial and harmful effects of ROS, the results suggest glucose-induced oxidative stress in Dictyostelium, a process that might originate from aerobic glycolysis, as well as a protective role of tetrahydropteridines against this stress.

Phosphoinositide turnover in Toll-like receptor signaling and trafficking

  • Le, Oanh Thi Tu;Nguyen, Tu Thi Ngoc;Lee, Sang Yoon
    • BMB Reports
    • /
    • v.47 no.7
    • /
    • pp.361-368
    • /
    • 2014
  • Lipid components in biological membranes are essential for maintaining cellular function. Phosphoinositides, the phosphorylated derivatives of phosphatidylinositol (PI), regulate many critical cell processes involving membrane signaling, trafficking, and reorganization. Multiple metabolic pathways including phosphoinositide kinases and phosphatases and phospholipases tightly control spatio-temporal concentration of membrane phosphoinositides. Metabolizing enzymes responsible for PI 4,5-bisphosphate (PI(4,5)P2) production or degradation play a regulatory role in Toll-like receptor (TLR) signaling and trafficking. These enzymes include PI 4-phosphate 5-kinase, phosphatase and tensin homolog, PI 3-kinase, and phospholipase C. PI(4,5)P2 mediates the interaction with target cytosolic proteins to induce their membrane translocation, regulate vesicular trafficking, and serve as a precursor for other signaling lipids. TLR activation is important for the innate immune response and is implicated in diverse pathophysiological disorders. TLR signaling is controlled by specific interactions with distinct signaling and sorting adaptors. Importantly, TLR signaling machinery is differentially formed depending on a specific membrane compartment during signaling cascades. Although detailed mechanisms remain to be fully clarified, phosphoinositide metabolism is promising for a better understanding of such spatio-temporal regulation of TLR signaling and trafficking.